Journal: Scientific Reports

Article Title: DEBIO 1143, an IAP inhibitor, reverses carboplatin resistance in ovarian cancer cells and triggers apoptotic or necroptotic cell death

doi: 10.1038/s41598-018-35860-z

Figure Lengend Snippet: In vitro inhibition of HOAC viability by carboplatin alone, or in combination with DEBIO 1143. HOAC (OVCAR-3, IGROV-1, A2780S, A2780R, SKOV-3, and EFO-21) were treated with increasing concentrations of carboplatin alone (from 1000 µM to 15.625 µM by 2-fold dilution steps) or in combination with DEBIO 1143 (10 µM) 24 h after seeding. ( A ) 48 h after treatment, cell viability was determined by a colorimetric assay using WST-1. The negative control (non-treated) of each condition corresponds to the 100% cell viability. ( B ) The IC50 of carboplatin after 48 h or 72 h of treatment was determined for each cell line (Mean +/− SEM, *p < 0.05, n = 3).

Article Snippet: The HOAC line EFO-21 was obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Germany).

Techniques: In Vitro, Inhibition, Colorimetric Assay, Negative Control

Journal: Scientific Reports

Article Title: DEBIO 1143, an IAP inhibitor, reverses carboplatin resistance in ovarian cancer cells and triggers apoptotic or necroptotic cell death

doi: 10.1038/s41598-018-35860-z

Figure Lengend Snippet: In vitro induction of apoptosis in HOAC by DEBIO 1143 alone or in combination with carboplatin. HOAC (OVCAR-3, SKOV-3, A2780S, and A2780R) were treated with carboplatin (IC50 after 48 h of treatment for each cell line), DEBIO 1143 (10 µM) or a combination of both treatments. The negative control corresponds to non-treated cells. ( A , B ) 48 h after treatment, cells were stained with a FITC-Annexin V/PI apoptosis detection kit. FITC-Annexin staining and PI incorporation were measured in cells with a FACS Calibur flow cytometer and analyzed with Cell Quest software. ( A ) Early apoptotic cells correspond to the Annexin V positive and PI negative population. (Mean +/− SEM, *p < 0.05, n = 3). ( B ) Late apoptotic and necrosis cells correspond to the Annexin V positive and PI positive population. (Mean +/− SEM, *p < 0.05, n = 3). ( C ) Western blots were used to analyze proteins extracts for the expression of PARP and cleaved PARP.

Article Snippet: The HOAC line EFO-21 was obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Germany).

Techniques: In Vitro, Negative Control, Staining, Flow Cytometry, Software, Western Blot, Expressing

Journal: Scientific Reports

Article Title: DEBIO 1143, an IAP inhibitor, reverses carboplatin resistance in ovarian cancer cells and triggers apoptotic or necroptotic cell death

doi: 10.1038/s41598-018-35860-z

Figure Lengend Snippet: In vitro blockade of apoptosis or necroptosis in HOAC treated with DEBIO 1143, alone or in combination with carboplatin. HOAC (SKOV-3 and EFO-21) were pre-treated for 2 h or not with Z-Vad 20 µM or necrostatin-1 50 µM, then treated with DEBIO 1143 (10 µM) or a combination of carboplatin (IC50 of each cell line after 48 h of treatment) and DEBIO 1143 (10 µM). 24 h after treatment, cell viability was determined by a colorimetric assay using WST-1. Cell viability inhibition percentage was determined compared to the negative control (no drug, NT) condition. (Mean +/− SEM, *p < 0.05, n = 3).

Article Snippet: The HOAC line EFO-21 was obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Germany).

Techniques: In Vitro, Colorimetric Assay, Inhibition, Negative Control

Journal: Scientific Reports

Article Title: DEBIO 1143, an IAP inhibitor, reverses carboplatin resistance in ovarian cancer cells and triggers apoptotic or necroptotic cell death

doi: 10.1038/s41598-018-35860-z

Figure Lengend Snippet: Effects of DEBIO 1143, alone or in combination with carboplatin, on the expression of targets. ( A ) HOAC (IGROV-1, SKOV-3, A2780S, A2780R, and EFO-21) were treated with carboplatin (IC 50 after 48 h of treatment for each cell line) ( C ), DEBIO 1143 (10 µM) ( D ) or a combination of both treatments (C+D). The negative control corresponds to non-treated cells (NT). Western blots were used to analyze proteins extracts for the expression of cIAP1 and XIAP. ( B ) cIAP1 and ( C ) XIAP proteins were quantified in 3 independent experiments and represented as relative quantities compared to the negative control. (Mean +/− SEM, *p < 0.05, **p < 0.01, n = 3).

Article Snippet: The HOAC line EFO-21 was obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Germany).

Techniques: Expressing, Negative Control, Western Blot

Journal: Scientific Reports

Article Title: DEBIO 1143, an IAP inhibitor, reverses carboplatin resistance in ovarian cancer cells and triggers apoptotic or necroptotic cell death

doi: 10.1038/s41598-018-35860-z

Figure Lengend Snippet: Role of caspase 9 expression in HOAC in response to DEBIO 1143. ( A ) Caspase 9 relative expression was determined in all cell lines by qPCR using GUSB, RPL19, TBP and TUBB as housekeeping genes (mean expression with EFO-21 caspase 9 expression as 1, *p < 0.05). ( B ) Protein extract from HOACs (A2780R, SKOV-3, and EFO-21 either control or following inhibition of caspase 9 expression) were analyzed by Western blots for the expression of caspase 9. The cells were treated with increasing concentrations of carboplatin alone (from 1000 µM to 15.625 µM by 2-fold dilution steps) or in combination with DEBIO 1143 (10 µM) 24 h after seeding. WT stands for wild-type. ( C ) 48 h after treatment, cell viability was determined by a colorimetric assay using WST-1. The negative control (non-treated) of each condition corresponds to the 100% cell viability. ( D ) The IC50 of carboplatin after 48 h of treatment was determined for each cell line (Mean +/− SEM, *p < 0.05, n = 4).

Article Snippet: The HOAC line EFO-21 was obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Germany).

Techniques: Expressing, Control, Inhibition, Western Blot, Colorimetric Assay, Negative Control